State Key Laboratory of Aridland Crop Science, Gansu Agricultural University, Lanzhou, 730070, China.
Hebei North University, Zhangjiakou, 075000, China.
BMC Genomics. 2024 Jan 2;25(1):10. doi: 10.1186/s12864-023-09890-5.
Plant U-box (PUB) E3 ubiquitin ligases have vital effects on various biological processes. Therefore, a comprehensive and systematic identification of the members of the U-box gene family in potato will help to understand the evolution and function of U-box E3 ubiquitin ligases in plants.
This work identified altogether 74 PUBs in the potato (StPUBs) and examined their gene structures, chromosomal distributions, and conserved motifs. There were seventy-four StPUB genes on ten chromosomes with diverse densities. As revealed by phylogenetic analysis on PUBs within potato, Arabidopsis, tomato (Solanum lycopersicum), cabbage (Brassica oleracea), rice (Oryza sativa), and corn (Zea mays), were clustered into eight subclasses (C1-C8). According to synteny analysis, there were 40 orthologous StPUB genes to Arabidopsis, 58 to tomato, 28 to cabbage, 7 to rice, and 8 to corn. In addition, RNA-seq data downloaded from PGSC were utilized to reveal StPUBs' abiotic stress responses and tissue-specific expression in the doubled-monoploid potato (DM). Inaddition, we performed RNA-seq on the 'Atlantic' (drought-sensitive cultivar, DS) and the 'Qingshu NO.9' (drought-tolerant cultivar, DT) in early flowering, full-blooming, along with flower-falling stages to detect genes that might be involved in response to drought stress. Finally, quantitative real-time PCR (qPCR) was carried out to analyze three candidate genes for their expression levels within 100 mM NaCl- and 10% PEG 6000 (w/v)-treated potato plantlets for a 24-h period. Furthermore, we analyzed the drought tolerance of StPUB25 transgenic plants and found that overexpression of StPUB25 significantly increased peroxidase (POD) activity, reduced ROS (reactive oxygen species) and MDA (malondialdehyde) accumulation compared with wild-type (WT) plants, and enhancing drought tolerance of the transgenic plants.
In this study, three candidate genes related to drought tolerance in potato were excavated, and the function of StPUB25 under drought stress was verified. These results should provide valuable information to understand the potato StPUB gene family and investigate the molecular mechanisms of StPUBs regulating potato drought tolerance.
植物 U-box(PUB)E3 泛素连接酶对各种生物过程有重要影响。因此,全面系统地鉴定马铃薯 U-box 基因家族的成员将有助于了解植物 U-box E3 泛素连接酶的进化和功能。
本研究共鉴定了马铃薯中的 74 个 PUB(StPUBs),并研究了它们的基因结构、染色体分布和保守基序。马铃薯的 74 个 StPUB 基因分布在 10 条染色体上,密度各不相同。通过对马铃薯、拟南芥、番茄(Solanum lycopersicum)、白菜(Brassica oleracea)、水稻(Oryza sativa)和玉米(Zea mays)中 PUB 的系统发育分析,它们被聚类为八个亚类(C1-C8)。根据与拟南芥、番茄、白菜、水稻和玉米的共线性分析,有 40 个 StPUB 基因与拟南芥、58 个与番茄、28 个与白菜、7 个与水稻、8 个与玉米同源。此外,还利用从 PGSC 下载的 RNA-seq 数据来揭示马铃薯加倍单倍体(DM)中 StPUB 对非生物胁迫的反应和组织特异性表达。此外,我们对开花早期、盛花期和落花期的‘大西洋’(耐旱品种,DS)和‘清薯 9 号’(耐旱品种,DT)进行了 RNA-seq,以检测可能参与响应干旱胁迫的基因。最后,进行了定量实时 PCR(qPCR)分析,以研究 100 mM NaCl 和 10%PEG 6000(w/v)处理的马铃薯幼苗中三个候选基因的表达水平,处理时间为 24 小时。此外,我们还分析了 StPUB25 转基因植株的耐旱性,发现与野生型(WT)植株相比,过表达 StPUB25 显著提高了过氧化物酶(POD)活性,降低了 ROS(活性氧)和 MDA(丙二醛)的积累,增强了转基因植株的耐旱性。
本研究挖掘了与马铃薯耐旱性相关的三个候选基因,并验证了 StPUB25 在干旱胁迫下的功能。这些结果应该为了解马铃薯 StPUB 基因家族和研究 StPUBs 调节马铃薯耐旱性的分子机制提供有价值的信息。